Cellular Antioxidant Properties of Human Natural Killer Enhancing Factor B

The protein, NKEF (natural killer enhancing factor), has been identified as a member of an antioxidant family of proteins capable of protecting against protein oxidation in cell-free assay systems. The mechanism of action for this family of proteins appears to involve scavenging or suppressing formation of protein thiyl radicals. In the present study we investigated the antioxidant protective properties of the NKEF-B protein overexpressed in an endothelial cell line (ECV304). Nkef-B-transfected cells displayed significantly lower levels of reactive oxygen species (ROS) compared with control or vector-transfected cells. Tert-Butylhydroperoxide-induced ROS was 15% lower in nkef-8-transfected cells and cytotoxicity was slightly, though not significantly, lower. NKEF-B had no effect on ROS induced by menadione or xanthine plus xanthine oxidase. NKEF-B overexpression resulted in slightly (≈ 10%) lower levels of cellular glutathione (GSH) and had no effect on rate or extent of GSH depletion following either diethylmaleate (DEM) or buthionine sulfoximine (BSO) treatment. Lipid peroxidation, assessed as thiobarbituric acid-reactive substances, was 40% lower in nkef-B-transfected cells compared with vector-only-transfected cells. DEM-induced lipid peroxidation was suppressed by NKEF-B at DEM concentrations of 20 μM to 1 mM. At 10 mM DEM, lipid peroxidation was unaffected by NKEF-B. NKEF-B expression also protected cells against menadione-induced inhibition of [³H]-thymidine uptake. The NKEF-B protein appears most effective in suppressing basal low-level oxidative injury such as that produced during normal metabolism. These results indicate that overexpression of the NKEF-B protein promotes resistance to oxidative stress in this endothelial cell line.     

Liquid chromatographic determination of gamma-aminobutyric acid in cerebrospinal fluid using 2-hydroxynaphthaldehyde as derivatizing reagent

gamma Aminobutyric acid (GABA) was determined by precolumn derivatization with 2-hydroxynaphthaldehyde and elution was made using Phenomenex C(18), 5 microm column with methanol: water (62:38 v/v) and UV detection at 330 nm. In a mixture containing glycine, l-lysine and tyramine GABA separated completely. A number of amines and amino acids tested did not affect the response of GABA. A linear calibration curve was obtained for GABA in the range of 1.2-28.0 microg/ml with detection limit of 2.8 ng/injection (5 microl). The method was used for the determination of GABA in cerebral spinal fluid (CSF) samples and gave results of 19.0 to 22.4 microg/m1 with coefficient of variation 2.4%     

An assessment of the relative effects of adverse physical and chemical properties of sodic soil on the growth and yield of wheat (Triticum aestivum L.)

Two wheat varieties were grown in artificially created sodic soils in pots at a range of sodicity levels (exchangeable sodium percentage (ESP) 15–52), with and without an anionic polyacrylamide soil conditioner (PAM) to stabilise soil aggregates. Increasing sodicity decreased the % water stable aggregates (% WSA) in soil and survival, grain and straw yield of wheat. Plants grown at high sodicity also had higher Na⁺, lower K⁺ and Ca²⁺ concentrations and lower K⁺/Na⁺ ratio in flag leaf sap than plants grown in control (non-sodic) soil. Sodicity had no effect on the concentrations of Cu²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ in grains and straw, but total uptake of these micronutrients was deceased due to lower dry weight of these tissues per plant. At all sodicity levels treatment of sodic soil with PAM increased the % WSA to values greater than in the non-sodic control soil, and slightly lowered ESP. Over the range ESP 15–44 the effects of PAM on wheat grain yield increased as sodicity increased, so that at ESP 44 grain yield in the treatment with PAM was only 25% lower than in the non-sodic control. However at ESP 52 the effects of PAM were smaller, and grain yield was 86% lower than in the control. At this sodicity level the decreases in grain yield due to sodicity and the increases in reponse to treatment of sodic soil with PAM were similar in the two varieties tested. At high sodicity levels (ESP 44 and 52) treatment of sodic soil with PAM decreased the concentration of Na⁺ and increased K⁺ and K⁺/Na⁺ ratio in flag leaf sap. However, at the highest sodicity level (ESP 52), flag leaf Na⁺ concentration remained above the level (100 mol m^-3) at which it has been found to be toxic. Concentrations of Cu²⁺, Fe²⁺, Mn²⁺ and Zn²⁺ in grain and straw were unaffected by PAM. These results suggest that at ESP up to 40–50 adverse physical characteristics are the major cause of low wheat yield in sodic soils, either due to their direct effects in decreasing growth, or their indirect effects in increasing uptake of Na⁺ and decreasing uptake of K⁺. Above ESP 50, roots are less able to exclude Na⁺, even in the presence of improved soil physical conditions, so that at these sodicity levels, both adverse physical and adverse chemical properties contribute to the decreased yield. This revised version was published online in June 2006 with corrections to the Cover Date.